OBJECTIVETo study the phylogeny relationship and molecular identification of 10 species from Huperzia (Huperziaceae) based on matK gene sequences data.

METHODTotal DNA of nine species from Huperzia was extracted; matK gene sequence was amplified by PCR. PCR product was directly sequenced after purification.

RESULTThe chloroplast matK gene nucleotide sequences from 9 species of Huperzia species were sequenced. The matK gene nucleotide sequences length was 1 589 bp. Analysis with Huperzia lucidula matK gene nucleotide sequences (download from GenBank) and taking Lycopodiella cernua as outgroup, Maximum Parsimony, Neighbor-Joining analyses and genetic distances were conducted using MEGA 3.1 software. 35 variable sites and 35 parsimony informative sites have been found. Pairwise genetic distances among 10 species of Huperzia was 1.59% - 0.25%.

CONCLUSIONThe results were consistent with the taxonomy in morphological of Huperzia. But H. longipetiolata and H. serrata were resolved into in different clade. There are 19 different sites of matK gene sequences between H. longipetiolata and H. serrata, the genetic distances is 0.121%. It is suggested H. longipetiolata should be as an independent species.

DNA, Chloroplast ; genetics ; DNA, Plant ; chemistry ; genetics ; Endoribonucleases ; genetics ; Huperzia ; classification ; enzymology ; genetics ; Molecular Sequence Data ; Nucleotidyltransferases ; genetics ; Phylogeny ; Plant Proteins ; genetics ; Plants, Medicinal ; classification ; enzymology ; genetics ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Species Specificity

ObjectiveTo explore the efficacy of constraint-induced movement therapy (CIMT) on motor function of upper extremity of chronic stroke patients.MethodsFifteen chronic stroke patients with hemiparesis (course of diseases more than 13.5 months) were treated by CIMT, involving restriction of movement of the intact upper extremity by placing it in a sling for 90% of waking hours for 12 days and training (by shaping) of the more affected extremity for 6 hours on the 10 workdays during that period. The therapeutic effect was evaluated with upper extremity function test (UEFT) and simple test for evaluating hand function (STEF).ResultsPatients showed a significant and very large degree of improvement after treatment on UEFT and STEF (ES, 0.8 and 0.5, respectively).ConclusionCIMT may be an efficacious method for improvement of the affected arm function of chronic stroke patients.

ObjectiveTo investigate the relationship between strength of the paretic lower limb and motor function, balance, walking speed, ability of daily living (ADL) in hemiparetic stroke patients.Methods85 stroke subjects, who were able to walk in the study, were evaluated in the strength of the paretic lower limb, motor function, balance, walking speed and ADL with Motricity Index, Fugl-Meyer Assessment, Berg Balance Scale, 10 m walking speed test and Functional Independence Measure (FIM). The levels of association between them were examined with Pearson's correlation coefficients and with multiple linear regression analyses by using the stepwise method. ResultsStrengths of the paretic lower limb were significantly positive related to motor function, balance, walking speed and ADL (r=0.592-0.811,P<0.001). The paretic ankle dorsiflexors, knee extensors, hip flexors were important clinical factor to consider in determining motor function(R2=0.377,P<0.001), balance(R2=0.321,P<0.001)and walking speed(R2=0.173,P<0.001), ADL(R2=0.42,P<0.001). ConclusionStrengths of the paretic lower limb of stroke patients may play an important role in their motor function, balance, walking speed and ADL.

OBJECTIVETo develop HDV as a vehicle to deliver hammerhead ribozyme into hepatocytes, the effects of modified HDV was assessed on the activity of embedded hammerhead ribozyme in vitro and in vivo.

METHODSIn vitro activity of ribozyme or HDV-driven ribozyme was assessed by incubating with the [alpha-32 P]-ATP labeled HBV RNA substrates at different temperature. Huh-7 cells were cotransfected with ribozyme or HDV-ribozyme chimera and HBV genome, by which inhibition of ribozymes on HBV transcription in vivo were examined.

RESULTSThe results indicated that both temperature and secondary structure influenced the cleavage activity of HDV-driven ribozyme significantly. When the factors were eliminated, the HDV-driven ribozyme could act as well as its counterpart naked ribozyme. While in cultured cells the HDV-driven ribozyme had higher inhibition to HBV gene expression than that of ribozyme alone.

CONCLUSIONThe results demonstrated that HDV may weaken the activity of embedded ribozyme in vitro, but make it enhanced in cultured cells. Thus, this study could provide a useful evidence to develop HDV as vector for liver-special delivery of ribozyme to against chronic HBV infection.

Base Sequence ; Cell Line, Tumor ; Genome, Viral ; Hepatitis Delta Virus ; enzymology ; genetics ; Humans ; Molecular Sequence Data ; Plasmids ; genetics ; Protein Structure, Secondary ; RNA, Catalytic ; chemistry ; genetics ; metabolism ; RNA, Viral ; genetics ; metabolism ; Structure-Activity Relationship ; Substrate Specificity ; Temperature ; Transfection

Animals ; Escherichia coli ; genetics ; metabolism ; Female ; Hepatitis B Core Antigens ; biosynthesis ; genetics ; immunology ; Hepatitis B Surface Antigens ; biosynthesis ; genetics ; immunology ; Hepatitis B Vaccines ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Protein Precursors ; biosynthesis ; genetics ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Vaccines, Synthetic ; immunology

OBJECTIVE: To establish 2-dimensional polyacrylamide gel electrophoresis (2-DE) map from human nasal polyps and normal nasal mucosa, and to identify differential expression proteins of 2-DE map. METHODS: Samples of nasal polyps and nasal mucosa (each sample group containing 7 cases) were obtained. The total proteins were extracted and separated by immobilized pH gradient (IPG)-based 2-DE. The silver-stained 2-DE was scanned with digital Imagescanner and analyzed with ImageMaster 2-DE Elite 4.01 software. To obtain peptide mass fingerprint (PMF) of differential protein spots, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was used. The PMF was searched in Swiss-Prot and TreMBL database by Pept-Ident software, to identify differential expression proteins. RESULTS: The well-resolved, reproducible 2-DE maps of nasal polyps and nasal mucosa were established. For the polyps tissues, the average proteins spot of three 2-DE maps was 825+/-78; and 682+/-96 spot was matched with the average matching rate of 82.7%. The average deviations of matched spot position were (1.13+/-0.16) mm in IEF direction and (1.45+/-0.21) mm in SDS-PAGE direction, respectively. For the nasal mucosa tissues, the average proteins spot of three 2-DE maps was 936+/-62; and 821+/-78 spots were matched with the average matching rate of 87.7%. After comparing the 2-DE maps of nasal polyps and nasal mucosa tissues, the protein spots were 1,458 and 1,617 respectively; and 1,026 protein spots were matched. Forty differential expression protein spots were incised from silver staining gel randomly and digested in the gel by TPCK-Trypsin. Thirty-four PMFs were obtained by MALDI-TOF-MS and 24 differential proteins were identified. CONCLUSION The well-resolved, reproducible 2-DE maps of human nasal polyps and nasal mucosa have been successfully established. Certain differential proteins related to the pathogenesis of human nasal polyps are identified.
Adult ; Electrophoresis, Gel, Two-Dimensional ; Female ; Humans ; Male ; Middle Aged ; Nasal Polyps ; metabolism ; Peptide Mapping ; Proteomics

ObjectiveTo determine the efficacy of constraint-induced movement therapy(CIMT) on different severity of the motor deficit of upper extremity after stroke.Methods27 stroke patients who had upper-limb hemiparesis and learned nonuse were allocated either to the moderate group(n=12) or the severe group(n=15) according to severity of initial motor deficit of upper extremity by Upper Extremity Function Test(UEFT).The two group patients were given CIMT,involving restriction of movement of the intact upper extremity by placing it in a sling for 90% of waking hours for 12 days and training(by repetitive practice,shaping) of the more affected extremity for 6 hours on the 10 weekdays during that period.Outcome were measured with UEFT.ResultsThere was no significant difference in treatment gain on the UEFT between the moderate patients and the severe patients(P>0.05).However,the Effect Size for the severe patients(ES=2.2) was larger than for the moderate patients(ES=1.8) at the quality of movement(UEFT).ConclusionCIMT is an effective rehabilitation technique for different severity of the motor deficit of upper extremity after stroke,especially in severe stroke patients.

OBJECTIVETo investigate the relationship between PPAR coactivator 1 (PGC-1), nuclear respiratory factor (NRF), mitochondrial transcription factor A (mtTFA) expressions of vascular smooth muscle cells (VSMC) and development of atherosclerosis in a rabbit model.

METHODSAtherosclerotic model was established by feeding the rabbits with high-fat diet for 4, 8 and 12 weeks (n = 10 each). Another 8 rabbits fed with normal diet served as normal controls. Intima-media ratio, mRNA and protein expressions of PGC-1, NRF, mtTFA and SMemb, a marker for synthetic VSMC, were detected on aorta specimens.

RESULTSWith the blood lipid increased, the intima-media ratio rose from (0.031 +/- 0.010) microm up to (0.814 +/- 0.258) microm during 12 weeks. Increasing SMemb means that synthetic VSMC grew more and more. The expressions of PGC-1 became significant after 4 weeks (P < 0.01), while that of NRF-1 and mtTFA rose significantly after 8 weeks (P < 0.01).

CONCLUSIONSThe PGC-NRF-mtTFA pathway might play a critical role in VSMC proliferation and development of atherosclerosis.

Animals ; Atherosclerosis ; blood ; metabolism ; pathology ; DNA-Binding Proteins ; metabolism ; Disease Models, Animal ; Female ; Lipids ; blood ; Male ; Mitochondrial Proteins ; metabolism ; Muscle, Smooth, Vascular ; metabolism ; Nuclear Respiratory Factor 1 ; metabolism ; Rabbits ; Trans-Activators ; metabolism ; Transcription Factors ; metabolism

BACKGROUNDTo investigate the immunological and virological efficacy of the therapeutic vaccine HBV CS1, a recombinant fusion protein which is composed of HBV core aa 1-155 plus PreS1 aa 3-55,against chronic HBV infection.

METHODSHBV transgenic mice were immunized with HBV CS1(5 ug) emulsified in equal volume of complete Freund adjuvant on day 0, followed by a second vaccination with HBV CS1(5 ug) emulsified with incomplete Freund adjuvant on days 21. Mice of control group were mock-vaccinated with PBS plus complete Freund adjuvant/incomplete Freund adjuvant. The splenocytes of individual mouse were subjected to T cell proliferation assays by using 3Hg thymidine, HBsAg and HBV DNA in sera of mice were detected by ELISA and quantitative PCR, respectively.

RESULTSHBV CS1 specific T cell response were induced in mice immunized with HBV CS1, with the titer of HBsAg and the level of HBV DNA decreased significantly after twice immunization with HBV CS1, while the control group almost remained the same.

CONCLUSIONHBV CS1 has the immunological and virological efficacy against chronic HBV infection in HBV transgenic mice; HBV CS1 could represent candidate vaccine for further studies on its role as therapeutic vaccine against HBV chronic infection in human.

Animals ; Female ; Hepatitis B Antibodies ; blood ; Hepatitis B Core Antigens ; genetics ; immunology ; therapeutic use ; Hepatitis B Surface Antigens ; genetics ; immunology ; therapeutic use ; Hepatitis B Vaccines ; genetics ; immunology ; therapeutic use ; Hepatitis B virus ; genetics ; immunology ; Hepatitis B, Chronic ; drug therapy ; immunology ; Humans ; Immunization ; Male ; Mice ; Mice, Transgenic ; Protein Precursors ; genetics ; immunology ; therapeutic use ; Random Allocation ; Recombinant Fusion Proteins ; genetics ; immunology ; therapeutic use

Objective To investigate the frequency of CD4~+CD25~+FoxP3~+regulatory T cells (Treg)and the expression of the functional protein,FoxP3,in patients with active tuberculosis and the relationship between Treg and the pathogenesis of tuberculosis.Methods Forty-five patients with active tuberculosis(including 25 cases of pulmonary tuberculosis and 20 tuberculous lymphadenitis), 20 healthy controls,20 recovered tuberculosis patients and 6 patients with reactive hyperplasia in cer- vical lymph node were enrolled.The frequency of CD4~+ CD25~+ FoxP3~+ Treg in the peripheral blood was measured by flow cytometry.FoxP3 mRNA expression was determined by real-time reverse transcriptase-polymerase chain reaction(RT-PCR)and the expression of FoxP3 protein in lymphoid tissues was measured by immunohistochemistry.Results The frequency of natural Treg in the peripheral blood from the patients with active tuberculosis was 2.91%?0.23%,which was signifi- cantly higher than that of healthy control group(1.22%?0.18%)and recovered tuberculosis patients(1.50%?0.17%,P